National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK.
Nat Commun. 2021 Apr 12;12(1):2168. doi: 10.1038/s41467-021-22376-w.
Quantifying small, rapidly progressing three-dimensional forces generated by cells remains a major challenge towards a more complete understanding of mechanobiology. Traction force microscopy is one of the most broadly applied force probing technologies but ascertaining three-dimensional information typically necessitates slow, multi-frame z-stack acquisition with limited sensitivity. Here, by performing traction force microscopy using fast single-frame astigmatic imaging coupled with total internal reflection fluorescence microscopy we improve the temporal resolution of three-dimensional mechanical force quantification up to 10-fold compared to its related super-resolution modalities. 2.5D astigmatic traction force microscopy (aTFM) thus enables live-cell force measurements approaching physiological sensitivity.
量化由细胞产生的微小、快速发展的三维力仍然是更全面地理解机械生物学的主要挑战。牵引力显微镜是应用最广泛的力探测技术之一，但确定三维信息通常需要使用有限灵敏度的缓慢、多帧 z 堆叠采集。在这里，通过使用快速单帧像散成像结合全内反射荧光显微镜进行牵引力显微镜测量，与相关的超分辨率模式相比，我们将三维机械力定量的时间分辨率提高了 10 倍。因此，2.5D 像散牵引力显微镜（aTFM）能够实现接近生理灵敏度的活细胞力测量。
